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Fluidigm Access Array - Picogreen Fluorimetry Quantification; Pool the Products from Multiple Access Array Ifcs

Fluidigm Access Array
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Access Array System for the Ion Torrent PGM Sequencing System: User Guide
Chapter 6: Post-PCR Amplicon Purification and Quantitation
PCR Product Library Quantitation Procedure
39
c The Region Table listed below the electropherogram will show the concentration of
the Region containing the PCR product library. Refer to the Agilent 2100 Bioanalyzer
User Guide for additional information on Regions.
PicoGreen Fluorimetry Quantification
Quantitate the PCR product library by fluorimetry, using the Quant-iT PicoGreen dsDNA
Assay Kit, following the manufacturer’s instructions.
Independent IFC libraries should be quantitated separately. After quantitation, libraries must
be normalized and volumetrically combined into a single final library for sequencing. The
final library concentration based on Quant-iT is then used for the downstream sequencing
preparation.
Pool the Products from Multiple Access Array IFCs
To pool amplicons from more than one Access Array IFC, pool and purify each Access Array
IFC separately.
1 Follow the steps outlined in this chapter under Purify Harvested PCR Products on
page 37 for each Access Array IFC.
2 For bidirectional amplicon tagged libraries (generated in Chapter 5), pool products A and
B from the same Access Array IFC (up to 48 samples) together for purification.

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